(1) Field of the Invention
The present invention relates to methods of amplification of nucleic acids. Specifically, nucleic acid amplification using duplex primers.
(2) Description of Related Art
There are a variety of methods for the amplification of nucleic acids known to those skilled in the art. Providing unique tail sequences during the amplification process can be useful for capturing, detecting or sequencing a target nucleic acid and eliminate the need to perform additional steps after amplification if labeling the target nucleic acid is desired. Furthermore, methods in the art that utilize random primers to assure good coverage of a number of target sequences in a sample also suffer from the problem that they can bind in a wide variety of locations on the target or target fragments, generating a wide range of amplicon sizes. The method disclosed in the present invention solves this problem by directing annealing of the random primers with tags more favorably to the ends of the target molecules and fragments thereof, as well as cDNA strands generated from these target molecules and fragments, thus generating longer and more uniform in size amplicons with tags incorporated at both ends. In addition, amplification methods often involve multiple steps. Reducing the number of steps required can save time and resources. The present invention provides a method for incorporating unique tail sequences during amplification as well as a duplex primer having a first strand for binding and expanding the target nucleic acid while the second strand may be used to expand the product.